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Characterization of Erysiphe necator-responsive genes in Chinese Wild Vitis quinquangularis.

Gao M, Niu J, Zhao S, Jiao C, Xu W, Fei Z, Wang X - Int J Mol Sci (2012)

Bottom Line: Genes involved in metabolism, photosynthesis, transport and signal transduction were also enriched in the library.Expression analysis of 13 selected genes by qRT-PCR revealed that most were induced more quickly and intensely in the resistant material "Shang-24" than in the sensitive V. pseudoreticulata clone "Hunan-1" by E. necator infection.The ESTs reported here provide new clues to understand the disease-resistance mechanism in Chinese wild grapevine species and may enable us to investigate E. necator-responsive genes involved in PM resistance in grapevine germplasm.

View Article: PubMed Central - PubMed

Affiliation: College of Horticulture, Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China, Ministry of Agriculture, Northwest A & F University, Yangling, Shaanxi 712100, China; E-Mails: gaomin.001@163.com (M.G.); niujiao.2009@163.com (J.N.); bashijiu@163.com (S.Z.); dd_race@sohu.com (C.J.) ; State Key Laboratory of Crop Stress Biology in Arid Areas, Northwest A & F University, Yangling, Shaanxi 712100, China.

ABSTRACT
Powdery mildew (PM), caused by fungus Erysiphe necator, is one of the most devastating diseases of grapevine. To better understand grapevine-PM interaction and provide candidate resources for grapevine breeding, a suppression subtractive hybridization (SSH) cDNA library was constructed from E. necator-infected leaves of a resistant Chinese wild Vitis quinquangularis clone "Shang-24". A total of 492 high quality expressed sequence tags (ESTs) were obtained and assembled into 266 unigenes. Gene ontology (GO) analysis indicated that 188 unigenes could be assigned with at least one GO term in the biological process category, and 176 in the molecular function category. Sequence analysis showed that a large number of these genes were homologous to those involved in defense responses. Genes involved in metabolism, photosynthesis, transport and signal transduction were also enriched in the library. Expression analysis of 13 selected genes by qRT-PCR revealed that most were induced more quickly and intensely in the resistant material "Shang-24" than in the sensitive V. pseudoreticulata clone "Hunan-1" by E. necator infection. The ESTs reported here provide new clues to understand the disease-resistance mechanism in Chinese wild grapevine species and may enable us to investigate E. necator-responsive genes involved in PM resistance in grapevine germplasm.

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Distribution of number of EST members in each grape unigene.
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f2-ijms-13-11497: Distribution of number of EST members in each grape unigene.

Mentions: To identify genes that are potentially involved in V. quinquangularis clone “Shang-24” resistance to E. necator, an SSH library was constructed with mock-inoculated “Shang-24” leaves as the driver and E. necator-inoculated leaves as the tester. A total of 605 clones from the SSH library were selected and PCR analysis of white E. coli colonies containing cDNA inserts in the pGEM-T Easy vector showed that the size of the cDNA inserts ranged from 150 to 1000 bp. This confirmed the quality of the subtracted cDNA library. A total of 526 clones were identified to contain inserts larger than 200 bp, and were selected for sequencing. After removing low quality sequences and sequences of bacterial origin, 492 high quality ESTs ranging between 200 and 1000 bp were obtained. These ESTs were further assembled into 266 unigenes, among which 101 were contigs and 165 were singletons (Table S1). The number of EST members in unigenes varied from one to 20 (Figure 2), with the majority of unigenes present in low copy numbers.


Characterization of Erysiphe necator-responsive genes in Chinese Wild Vitis quinquangularis.

Gao M, Niu J, Zhao S, Jiao C, Xu W, Fei Z, Wang X - Int J Mol Sci (2012)

Distribution of number of EST members in each grape unigene.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC3472759&req=5

f2-ijms-13-11497: Distribution of number of EST members in each grape unigene.
Mentions: To identify genes that are potentially involved in V. quinquangularis clone “Shang-24” resistance to E. necator, an SSH library was constructed with mock-inoculated “Shang-24” leaves as the driver and E. necator-inoculated leaves as the tester. A total of 605 clones from the SSH library were selected and PCR analysis of white E. coli colonies containing cDNA inserts in the pGEM-T Easy vector showed that the size of the cDNA inserts ranged from 150 to 1000 bp. This confirmed the quality of the subtracted cDNA library. A total of 526 clones were identified to contain inserts larger than 200 bp, and were selected for sequencing. After removing low quality sequences and sequences of bacterial origin, 492 high quality ESTs ranging between 200 and 1000 bp were obtained. These ESTs were further assembled into 266 unigenes, among which 101 were contigs and 165 were singletons (Table S1). The number of EST members in unigenes varied from one to 20 (Figure 2), with the majority of unigenes present in low copy numbers.

Bottom Line: Genes involved in metabolism, photosynthesis, transport and signal transduction were also enriched in the library.Expression analysis of 13 selected genes by qRT-PCR revealed that most were induced more quickly and intensely in the resistant material "Shang-24" than in the sensitive V. pseudoreticulata clone "Hunan-1" by E. necator infection.The ESTs reported here provide new clues to understand the disease-resistance mechanism in Chinese wild grapevine species and may enable us to investigate E. necator-responsive genes involved in PM resistance in grapevine germplasm.

View Article: PubMed Central - PubMed

Affiliation: College of Horticulture, Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China, Ministry of Agriculture, Northwest A & F University, Yangling, Shaanxi 712100, China; E-Mails: gaomin.001@163.com (M.G.); niujiao.2009@163.com (J.N.); bashijiu@163.com (S.Z.); dd_race@sohu.com (C.J.) ; State Key Laboratory of Crop Stress Biology in Arid Areas, Northwest A & F University, Yangling, Shaanxi 712100, China.

ABSTRACT
Powdery mildew (PM), caused by fungus Erysiphe necator, is one of the most devastating diseases of grapevine. To better understand grapevine-PM interaction and provide candidate resources for grapevine breeding, a suppression subtractive hybridization (SSH) cDNA library was constructed from E. necator-infected leaves of a resistant Chinese wild Vitis quinquangularis clone "Shang-24". A total of 492 high quality expressed sequence tags (ESTs) were obtained and assembled into 266 unigenes. Gene ontology (GO) analysis indicated that 188 unigenes could be assigned with at least one GO term in the biological process category, and 176 in the molecular function category. Sequence analysis showed that a large number of these genes were homologous to those involved in defense responses. Genes involved in metabolism, photosynthesis, transport and signal transduction were also enriched in the library. Expression analysis of 13 selected genes by qRT-PCR revealed that most were induced more quickly and intensely in the resistant material "Shang-24" than in the sensitive V. pseudoreticulata clone "Hunan-1" by E. necator infection. The ESTs reported here provide new clues to understand the disease-resistance mechanism in Chinese wild grapevine species and may enable us to investigate E. necator-responsive genes involved in PM resistance in grapevine germplasm.

Show MeSH
Related in: MedlinePlus